The objective of this project are to obtain an understanding of the nature and function of membranes. Oxidative phosphorylation, electron transport and active transport of amino acids, are all membrane related phenomena which require specific spatial orientation of the respiratory carriers and transport binding-carrier proteins in order to function. Studies will be continued in the elucidation of the carriers involved, their topology or location in or on the membrane, as well as the underlying driving force involved. The mechanism, role and regulation of Ca2 ion transport will be studied with a cell-free bacterial system. Particular attention will be paid to the mechanism and role of proteins which contain gamma-carboxyglutamic acid residues. Further insight into the bioenergetic mechanism involved with active transport of amino acids may be obtained from the model proteoliposomal system containing the purified "binding"-carrier protein for calcium transport. The physicochemical properties of the purified carrier protein will also be studied. In addition, studies will be undertaken to determine the nature and sequences of the respiratory carriers in order to fully characterize the respiratory chain in M. phlei. These studies of membrane-related phenomena will provide insight into the broader questions of the nature, topology of membrane proteins, conformational changes and functional aspects of membranes. Studies will also be undertaken to understand the role and specificity of quinones in the respiratory chain as well as their possible participation in formation of a proton gradient, membrane potential, Ca2 ion transport and oxidative phosphorylation.